Introducing: ONI’s Next-Generation End-to-End Solution for EV Research Learn More
Harness the power of super-resolution microscopy to characterize EV populations and individual EVs. Quantify biomarkers on small, rare, and precious EVs with single molecule sensitivity. ONI’s EV Profiler helps you focus on answering questions about your EVs.
With the EV Profiler Kit you use our validated antibodies to measure the expression of tetraspanins and characterize the single, double and triple positive populations, or you can measure your own biomarker of interest.*
Single molecule localization microscopy used in combination with the appropriate fluorescently labeled probes, helps confirm EV identity after purification. ONI’s EV Profiler Kit in combination with cluster-based analysis on our CODI cloud platform, allows reporting of morphological features of the EV, including size and circularity.
*Reagents have a shelf life of two months.
ONI’s EV Profiler Kit allows detection of up to 3 biomarkers on captured EVs. HCT116-derived EV sample were captured with ONI’s S4 molecule, labeled with anti-CD9-CF®488 (yellow), anti-CD63-CF®568 (cyan) and anti-CD81-CF®647 (magenta) and imaged using dSTORM on the Nanoimager (left). Biomarker positivity was quantified using ONI’s CODI cluster-based algorithm on single-molecule localization microscopy data (right).
EV Profiler enables the capture of EVs on a passivated glass surface. The kit’s workflow is compatible with the following capture methods:
Validated by ONI – to be selected during purchase:
Choose your own:
ONI validated capture antibodies against tetraspanins: anti-CD9, anti-CD63 and anti-CD81 antibodies can be selected when purchasing the kit.
Our proprietary S4 capture molecule binds EVs in a protein-independent manner, working best with phosphatidylserine-rich EVs.
Use your own antibody of choice against your target of interest – it simply needs to be biotinylated (sample above provided by Dr Nicole Meisner-Kober’s lab).
Study EV populations across scales. Each EV Profiler kit contains enough material to analyze 12 EV samples using our 4 lane analysis chip which has been developed and optimized for EV capture. You can capture and image hundreds of EVs from any given source with the appropriate capture method (read more details below), then zoom into individual EVs to see biomarker distribution at a single-EV level.
ONI’s Nanoimager and CODI platform enable imaging and visual inspection of EV samples across scales. Scale bars range from 10 µm to 50 nm when inspecting one EV out of thousands of EVs imaged on from an HCT116-derived EV sample captured with ONI’s S4 molecule and labeled with pan-EV stain (yellow), anti-CD63-CF®568 (cyan) and anti-CD81-CF®647 (magenta).
The EV Profiler Kit contains all necessary reagents to capture, fix and stain EVs ready for imaging and the workflow works with EV sample volumes as low as 10 µL.
Single molecule localization data and nanoscale resolution images can be obtained using the Nanoimager, our benchtop super-resolution microscope. Data is then analyzed through our CODI cloud-based software, making it possible to get results within 24 hours from start to finish.
Besides the flexibility that EV Profiler offers for different EV capture methods, you can easily swap the fluorescent antibodies for surface marker detection for your own.
In the EV Profiler Kit, antibodies for standard tetraspanin markers (CD9, CD63, and CD81) are included conjugated to three different fluorophores so you can profile these markers. However, the kit is modular and antibodies can be swapped for your own validated antibodies against targets of interest inside or outside EVs. These need to be conjugated to dSTORM-compatible dyes.
You can find more information on fluorophores for dSTORM imaging here. The kit’s sample preparation protocol will guide you through the steps for incorporating your antibodies into the experiment and our image acquisition and analysis guide will provide you with the information needed when optimizing your antibodies.
Purchase of the kit includes access to purpose built analysis tools within our CODI cloud platform, which allow seamless processing of single molecule localization data to obtain output summary data files.
ONI’s CODI cloud software uses clustering of single molecule data to enable study of EVs. Left image shows EVs captured using the EV Profiler Kit, which were imaged with the Nanoimager. Single molecule localization data is then clustered on CODI to help assess EV morphology features and determine levels of positivity for different biomarkers.
Combine the advantages of super-resolution imaging with powerful analysis of larger numbers of events. This allows detection and quantification of biomarker abundance with up to 3 biomarkers detected within the same sample using dSTORM, and reporting of EV morphology, including size and circularity.
Chips |
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| 3 four lane flow-cell chips | |
| 6 lane stickers | |
| 2 stage securing magnets | |
Capture reagents (3 to choose from) |
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| Phosphatidylserine-dependent S4 capture | |
| Biotinylated anti-CD9 capture | |
| Biotinylated anti-CD63 capture | |
| Biotinylated anti-CD81 capture | |
Detection probes |
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| Anti-CD9 488 (1 tube) | |
| Anti-CD63 568 (1 tube) | |
| Anti-CD81 647 (1 tube) | |
Standards |
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| E1 standard EVs (1 tube) | |
Buffers |
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| S3, W1, F1, B1 (3 tubes each) | |
| C1 (1 tube) | |
| BCubed imaging buffer parts A and B (3 tubes each) | |
